Matrix droplet extruder, sample holder and sample analysis system

ABSTRACT

A matrix droplet extruder includes a casing; one or a plurality of reagent containers; a plurality of pneumatic connectors on the casing that are each configured to be connected to a pneumatic actuator to provide controlled pneumatic pressure to one or more of said plurality of the pneumatic connectors; a droplet matrix extrusion surface with one or a plurality of printing zones, each printing zone comprising an array of perforations; and a liquid management chip for dispensing one or more reagents from said one or a plurality of reagent containers through the array of perforations of said one or a plurality of printing zones, so as to repeatedly generate a matrix of droplets when applying the pneumatic pressure to said one or more reagents.

FIELD OF THE INVENTION

The present invention relates to a sample analysis system and method.

BACKGROUND OF THE INVENTION

In many cases, rapid receipt of test results is important for a timelymedical diagnosis. Different approaches have been used to facilitateattaining of timely results. Large, automated laboratories may enablerapid testing of large numbers of samples. However, a test sample (e.g.,of blood, saliva, or another tested material) must be transported to thelaboratory from a local clinic or other facility. Point-of-care (POC)testing generally refers to diagnostic tests that are performed outsideof a central laboratory, e.g., at a health care clinic or other facilitywhere a patient is present. Over the years, the increasing availabilityof transportable, portable, and, in some cases, handheld instrumentshave resulted in the migration of POC testing from the hospitalenvironment to a range of medical environments including the workplace,home, disaster care facilities, and local clinics.

Use of POC testing affects many types of diagnostics, includinginfectious diseases, autoimmune conditions, allergies, cancer, cardiacdisease, bowel diseases, and more.

A typical POC testing device provides a single diagnostic test for asingle sample. However, in many scenarios, a panel of diagnostic testsis required. Therefore, use of such single-test typical POC testingdevices may require a clinic to acquire and maintain a wide variety ofdifferent POC devices.

SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described andillustrated in conjunction with systems, tools and methods which aremeant to be exemplary and illustrative, not limiting in scope.

There is thus provided, in accordance with an embodiment of theinvention, a matrix droplet extruder. The matrix droplet extruder mayinclude a casing; one or a plurality of reagent containers; a pluralityof pneumatic connectors on the casing that are each configured to beconnected to a pneumatic actuator to provide controlled pneumaticpressure to one or more of said plurality of the pneumatic connectors.The matrix droplet extruder may also include a droplet matrix extrusionsurface with one or a plurality of printing zones, each printing zonecomprising an array of perforations; and a liquid management chip fordispensing one or more reagents from said one or a plurality of reagentcontainers through the array of perforations of said one or a pluralityof printing zones, so as to repeatedly generate a matrix of dropletswhen applying the pneumatic pressure to said one or more reagents.

In some embodiments of the present invention, the droplet matrixextrusion surface comprises a pattern for allowing mixing of two or moredroplets.

In some embodiments of the present invention, the matrix dropletextruder may further include a washer for washing said one or aplurality of printing zones.

In some embodiments of the present invention, the washer is configuredto move between an idle stand-by position away from said one or aplurality of printing zones and a washing position over said one or aplurality of printing zones.

In some embodiments of the present invention, the washer comprises aninlet port for introducing and for evacuating a washing agent onto andfrom said one or a plurality of printing zones.

In some embodiments of the present invention, the matrix dropletextruder may further include a pneumatic pressure actuator; a controllerto control the pneumatic pressure actuator; and a docking station havinga plurality of pneumatic docking station connectors connected to thepneumatic pressure actuator.

In some embodiments of the present invention, the controller isconfigured to control a period of time during which pressure is appliedby the pneumatic pressure actuator to a pneumatic connector of theplurality of pneumatic connectors.

In some embodiments of the present invention, the controller isconfigured to control a pressure that is applied by the pneumaticpressure actuator to a pneumatic connector of the plurality of pneumaticconnectors.

In some embodiments of the present invention, a sample holder mayinclude a casing with a pocket; a removable sample slide with one or aplurality of substrate patches configured to be inserted into or removedfrom the pocket; a sample channel for introducing a liquid sample intoone or a plurality of sealed spaces between the sample slide and asection of the casing, so as to allow the sample liquid to be absorbedin said one or more substrate patches.

In some embodiments of the present invention, the substrate patchescomprise a membrane.

In some embodiments of the present invention, the membrane includesmaterial selected from the group of materials consisting of:nitrocellulose, fiberglass, nano-mesh, plastic and glass.

In some embodiments of the present invention, the sample holder includesa plurality of pneumatic connectors on the casing that are eachconfigured to be connected to a pneumatic pressure or vacuum actuator toprovide controlled pneumatic pressure or vacuum to one or more of saidplurality of the pneumatic connectors to obtain laminar flow of thesample or other liquids into said one or a plurality of sealed spaces.

In some embodiments of the present invention, the sample holder includesone or a plurality of liquid containers containing one or more kinds ofthe other liquids.

In some embodiments of the present invention, the sample holder includesa lock for locking the sample slide when fully inserted inside thepocket.

In some embodiments of the present invention, there is provided a sampleanalysis system that includes a pneumatic actuator; one or a pluralityof sample holders comprising: a casing with a pocket; liquid containers;a removable sample slide with a substrate comprising one or a pluralityof substrate patches, configured to be inserted into or removed from thepocket; and a sample channel for introducing a liquid sample into one ora plurality of sealed spaces between the sample slide and a section ofthe casing, so as to allow the sample liquid to be absorbed in said oneor more substrate patches; a controller that is configured to controlthe pneumatic actuator so as to sequentially cause one or more liquidsfrom one or more of the liquid containers to flow to or away from thesample slide in that sample holder; and an analysis module for examiningthe sample slide after contact of the sample with a plurality of dots ofone or more reagents that are printed onto the substrate.

In some embodiments of the present invention, the analysis system alsoincludes a transport mechanism to transport each of said one or aplurality of the sample holders sequentially to a station of a pluralityof stations along the transport mechanism, the controller furtherconfigured to control the transport mechanism so as to cause thetransport mechanism to move said one or a plurality of sample holderssuch that each of the one or plurality of sample holders is sequentiallymoved from a current station of the plurality of stations to a nextstation of the plurality of stations.

In some embodiments of the present invention, the transport mechanismcomprises a rotatable carousel, the plurality of pneumatic connectorsarranged to enable connection of each of said one or a plurality ofsample holders at a position on the perimeter of the carousel.

In some embodiments of the present invention, the plurality of pneumaticconnectors are arranged to enable connection of said one or a pluralityof sample holders at equally spaced positions along the perimeter of thecarousel.

In some embodiments of the present invention, the plurality of stationscomprises at least one docking station onto which a matrix dropletextruder is attachable, the docking station comprising a plurality ofdocking station connectors for connecting to a plurality of dockingpneumatic connectors of the matrix droplet extruder, the docking stationbeing controllable by the controller to selectively apply pressure toeach of the docking station connectors to control extrusion of liquiddroplets from one or more reagent containers of the matrix dropletextruder onto a droplet matrix extrusion surface of the matrix dropletextruder to form the plurality of reagent droplets.

In some embodiments of the present invention, said at least one dockingstation comprises a mechanism for emptying a canister of a reagent intoa reagent container of said one or more reagent containers.

In some embodiments of the present invention, the analysis systemincludes a mechanical system that is controlled by the controller tomanipulate the substrate of each of the one or plurality of sampleholders to place the substrate in contact with the droplet matrixextrusion surface to enable extrusion of the plurality of reagent dotsonto the substrate, and to expose the substrate to the analysis module.

In some embodiments of the present invention, the mechanical system iscontrolled by pneumatic pressure.

In some embodiments of the present invention, the mechanical systemcomprises a rinsing chamber that is configured to be placed against thedroplet matrix extrusion surface to enable cleaning of the dropletmatrix extrusion surface.

In some embodiments of the present invention, the analysis modulecomprises an optical system for acquiring an image of the substrate oran electronic system to measure an electrical property of the substrate.

In some embodiments of the present invention, the substrate includessaid dots of one or more reagents which are pre-extruded onto thesubstrate.

In some embodiments of the present invention, the analysis systemfurther includes a matrix droplet extruder for printing the plurality ofdots of one or more reagents. The matrix droplet extruder includes acasing; one or a plurality of reagent containers; a second plurality ofpneumatic connectors on the casing that are each configured to beconnected to the pneumatic actuator to provide controlled pneumaticpressure to one or more of said second plurality of the pneumaticconnectors; a droplet matrix extrusion surface with one or a pluralityof printing zones, each printing zone comprising an array ofperforations; and a liquid management chip for dispensing one or morereagents from said one or a plurality of reagent containers through thearray of perforations of said one or a plurality of printing zones, soas to repeatedly generate a matrix of droplets when applying thepneumatic pressure to said one or more reagents.

In some embodiments of the present invention said one or a plurality ofprinting zones comprises two printing zones.

In some embodiments of the present invention, the analysis systemfurther includes a transfer mechanism for transferring each of said oneor a plurality of sample holders to and from a position facing thematrix droplet extruder, and for placing the sample slide of each ofsaid one or a plurality of sample holders onto either of said one or aplurality of printing zones.

In some embodiments of the present invention, the analysis systemfurther includes a disposal container for disposing used sample holdersof said one or a plurality of sample holders.

BRIEF DESCRIPTION OF THE DRAWINGS

In order for the presented invention, to be better understood and forits practical applications to be appreciated, the following Figures areprovided and referenced hereafter. It should be noted that the Figuresare given as examples only and in no way limit the scope of theinvention. Like components are denoted by like reference numerals.

FIG. 1 schematically illustrates a multiple sample analysis device, inaccordance with a possible embodiment of the present invention;

FIG. 2 is a block diagram of a multiple sample analysis device generallysimilar to that shown in FIG. 1;

FIG. 3A schematically illustrates an example of a transport and analysissystem of a multiple sample analysis device, possibly similar to thatshown in FIG. 2.

FIG. 3B is a schematic exploded view of the transport and analysissystem shown in FIG. 3A;

FIG. 4 is a schematic top view of the transport and analysis systemshown in FIG. 3A;

FIG. 5 schematically illustrates an example of a carousel of thetransport and analysis system shown in FIG. 3;

FIG. 6A schematically illustrates an example of a sample holder of amultiple sample analysis device such as that shown in FIG. 3;

FIG. 6B schematically illustrates a chamber for applying suction to aslide in the sample holder shown in FIG. 6A;

FIG. 7A schematically illustrates an example of a matrix dropletextruder inserted into an example of a docking station, possiblysuitable for use with a transport and analysis system such as that shownin FIG. 3A;

FIG. 7B is a schematically illustrates the docking station shown in FIG.7A;

FIG. 7C schematically illustrates a rear side of a matrix dropletextruder, such as that shown in FIG. 7A;

FIG. 7D is a schematic cross section of an example of a matrix dropletextruder such as that shown in FIG. 7C;

FIG. 7E schematically illustrates an example of a slide onto whichreagent dots were extruded, possibly by a matrix droplet extruder suchas that shown in FIG. 7A;

FIG. 8 schematically illustrates possible stations for sample holders ofan example of a transport and analysis system such as that shown in FIG.3A;

FIG. 9 is a flowchart depicting a possible method of operation of amultiple sample analysis device, in accordance with an embodiment of thepresent invention;

FIG. 10 schematically illustrates an example of a reagent dot printingsystem; and

FIG. 11 schematically illustrates an example of a standalone sampleprocessing system.

FIG. 12A is a front view of a sample holder 1200 of the multiple sampleanalysis device, according to some embodiments of the present invention,with a sample slide inserted inside.

FIG. 12B is a front view of a sample holder 1200 of the multiple sampleanalysis device, according to some embodiments of the present invention,with the sample slide pulled out.

FIG. 13A shows a matrix droplet extruder configured to perform dualprinting assays, according to some embodiments of the presentapplication, with a sample slide in a first print position.

FIG. 13B shows a matrix droplet extruder configured to perform dualprinting assays, according to some embodiments of the presentapplication, with a sample slide in a second print position.

FIG. 13C shows the back of a matrix droplet extruder, according to someembodiments of the present invention.

FIG. 13D is a front view of a washer of the matrix droplet extrudershown in FIG. 13A and FIG. 13B.

FIG. 14 is a schematic illustration of a sample analysis system,according to some embodiments of the present invention, with dualprinting assay capability.

It will be appreciated that for simplicity and clarity of illustration,elements shown in the figures have not necessarily been drawn to scale.For example, the dimensions of some of the elements may be exaggeratedrelative to other elements for clarity. Further, where consideredappropriate, reference numerals may be repeated within the figures toindicate like elements.

DETAILED DESCRIPTION OF THE INVENTION

In the following detailed description, numerous specific details are setforth in order to provide a thorough understanding of the invention.However, it will be understood by those of ordinary skill in the artthat the invention may be practiced without these specific details. Inother instances, well-known methods, procedures, components, modules,units and/or circuits have not been described in detail so as not toobscure the invention.

Although embodiments of the invention are not limited in this regard,discussions utilizing terms such as, for example, “processing,”“computing,” “calculating,” “determining,” “establishing”, “analyzing”,“checking”, or the like, may refer to operation(s) and/or process(es) ofa computer, a computing platform, a computing system, or otherelectronic computing device, that manipulates and/or transforms datarepresented as physical (e.g., electronic) quantities within thecomputer's registers and/or memories into other data similarlyrepresented as physical quantities within the computer's registersand/or memories or other information non-transitory storage medium(e.g., a memory) that may store instructions to perform operationsand/or processes. Although embodiments of the invention are not limitedin this regard, the terms “plurality” and “a plurality” as used hereinmay include, for example, “multiple” or “two or more”. The terms“plurality” or “a plurality” may be used throughout the specification todescribe two or more components, devices, elements, units, parameters,or the like. Unless explicitly stated, the method embodiments describedherein are not constrained to a particular order or sequence.Additionally, some of the described method embodiments or elementsthereof can occur or be performed simultaneously, at the same point intime, or concurrently. Unless otherwise indicated, the conjunction “or”as used herein is to be understood as inclusive (any or all of thestated options).

Some embodiments of the invention may include an article such as acomputer or processor readable medium, or a computer or processornon-transitory storage medium, such as for example a memory, a diskdrive, or a USB flash memory, encoding, including or storinginstructions, e.g., computer-executable instructions, which whenexecuted by a processor or controller, carry out methods disclosedherein.

In accordance with at least certain embodiments of the presentinvention, an analysis or diagnostic system (e.g., a point-of-caresystem) may be configured to enable non-automated or automatedsequential chemical, biochemical, and/or other analysis and/or testingof one of more (e.g. multiple) samples. For example, a sample mayinclude a substance derived from a person or animal (e.g., blood,saliva, urine, feces, or other type of material drawn from, ordischarged or secreted by, a human or animal patient), from anenvironment (e.g., water or other liquids, precipitation, mud or soil, amaterial extracted from or secreted by a plant, or other materials foundin an environment), from industrial products (foodstuffs, fuels,lubricants, pharmaceuticals, cleaning and maintenance products, beautyproducts, materials produced for industrial product, or other industrialproducts), and/or other types of substances or refined liquids.

It should be noted that reference is made herein to POC applications,systems, devices, and methods by way of example only. A system, device,or method as described herein may be used or applied in settings otherthan a medical clinic or similar setting. For example, devices, systems,and methods as described herein may be used in central laboratories, ina factory (e.g., for industrial or manufacturing applications), in thefield (e.g., for environmental, geological, biological, resourceexploration, or other applications), in a police laboratory (e.g., forforensic applications), or elsewhere. Reference herein to point-of-careshould be understood, therefore, as being equally applicable to anyother setting or application.

In order that a testing system may function effectively as apoint-of-care diagnostics system, the system may be arranged follow anapproved standard analysis protocol, e.g., meet Clinical LaboratoryImprovement Amendments (CLIA) standards, enzyme-linked immunosorbentassay (ELISA) protocol, DNA/RNA testing protocols, or another approvedprotocol, as well as meeting standards for prevention of mixing ofsamples from different patients.

In addition, the system may be configured so as to prevent misuse by theuser (e.g., improper insertion of a sample or improper application of atest to a sample). A test that may be performed by a system may notnecessarily require an excessive amount of time (e.g., no more than 10minutes to 20 minutes). In many cases, a qualitative (e.g., yes/no,positive/negative, present/not present, or similar) response may besufficient and may be preferable to a quantitative response in somecases.

Each sample may be spread onto a substrate, such as a membrane, on apossible slide of a separate sample holder. The sample holder may be asingle use device that may be discarded or disposed of after applicationof a test protocol to the sample.

For example, the sample holder may include one or more openings toenable introduction of a liquid sample into the sample holder. Forexample, each opening may open into an internal capillary for drawing aliquid sample, e.g., a drop of blood, or another liquid sample, into thesample holder. A user, e.g., healthcare professional or laboratorytechnician, a patient trained for self-testing, or other user, mayidentify the sample holder, e.g., by scanning a barcode, radiofrequencyidentification (RFID) tag, or otherwise, and may be matched with theinserted sample (e.g., patient identifier). The user may also input intothe system, or otherwise specify, a test protocol for application to aparticular sample in the identified sample holder.

The sample holder may be configured to apply to the sample a processingprotocol of one or more steps. Typically, the protocol may includeplacing the sample into contact with a droplet matrix extrusion surfaceof a matrix droplet extruder. For example, the slide may be mechanicallyremoved from the sample holder and pressed against the droplet matrixextrusion surface.

When the droplet matrix extrusion surface is in contact with the sample,or prior to contact, the matrix droplet extruder extrudes a matrix ofreagent dots onto the droplet matrix extrusion surface via acorresponding array of perforations that open out of the matrix dropletextruder at the droplet matrix extrusion surface. The selection ofreagents in each matrix of reagent droplets that is extruded may bespecific to a particular test protocol. The matrix droplet extruder maybe configured to include a plurality of liquid reservoirs.

The matrix droplet extruder may include a microfluidic channeling systemthat may be controllable to channel a liquid from one or more of theliquid reservoirs to a particular opening of an array of micro-openingsor perforations in the droplet matrix extrusion surface. The channeledliquid may be extruded via the opening onto a predetermined position onthe droplet matrix extrusion surface to form a reagent dot that isbrought into contact with the sample on the slide substrate of thesample holder.

The processing protocol may typically be configured to result in anoptically or electronically detectable result that is indicative of theresults of application of the analysis protocol (e.g., presence of aparticular substance, such as an antigen, in the sample). For example,the analysis protocol may be configured to change a color orfluorescence of a region of the sample slide that comes into contactwith one or more of the reagent dots, the color being indicative of thecomposition of the sample. Alternatively or in addition, the region ofthe sample slide that comes into contact with one or more of the reagentdots may fluoresce with a particular color that is indicative of thecomposition of the sample.

Alternatively or in addition, the region of the sample slide that comesinto contact with one or more of the reagent dots may be characterizedby one or more changes in electrical properties that are indicative ofthe composition of the sample. For example, the processing protocol mayinclude an immunoassay protocol, e.g., an enzyme-linked immunosorbentassay (ELISA) protocol, in which a sample is exposed to one or moreantibodies or enzymes in order to detect the presence of one or moreantigens in the sample.

In at least some examples, a multiple sample diagnostic system may beprovided that may include an integrated transport and control mechanismfor transporting several sample holders, each to one or more of aplurality of stations. The transport and control mechanism may beconfigured to concurrently transfer all of the sample holders from onestation to a next station, in succession. For example, the transportmechanism may include a linear or circular conveyor or carousel. In acase of circular carousel, the sample holders may be arranged along aperimeter of the carousel. In some cases, such a carousel may beconfigured to concurrently transport a maximum of eight sample holders(e.g., corresponding to a maximum number of processing steps in typicalELISA protocols), or another number of sample holders.

Each sample holder may include a plurality of pneumatic connectors thatmay be arranged to connect with corresponding connectors on thetransport mechanism when the sample holder is inserted into thetransport mechanism. The connectors may typically be connected to apressurized pneumatic source or to a vacuum pump or other suction sourcevia an arrangement of pneumatic channels and valves. Each sample holdermay also include one of more filters, reservoirs of one or morematerials, typically liquids, and/or other structure(s).

When the sample holder is positioned at some of the stations, the valvesmay be operated to cause movement of a quantity of liquid within thesample holder. For example, a sample of blood may be drawn through afilter arrangement in order to obtain blood plasma or serum that issuitable for testing. A liquid may be drawn from one or more of thereservoirs through to the sample on the substrate of the slide toperform a wet process procedure of a protocol (e.g., wash, blocking, orother step).

The transport mechanism may be configured such that each sample holderremains at each station for a period of time that is sufficient toenable completion of a step of a processing protocol on each of thesamples. For example, in many cases, a period of one minute may besufficient to perform the longest steps of typical analysis protocols.

The diagnostic system may include one or more (e.g., two) dockingstations to each of which a matrix droplet extruder may be connected(e.g., at the beginning of a work day, or otherwise). When, duringoperation of some protocols, a sample holder is brought to a processingstation that includes a matrix droplet extruder, the sample substrate ofthat sample holder may be pressed (e.g., after possible removal of thesample slide from the sample holder) against the droplet matrixextrusion surface of that matrix droplet extruder. Thus, the sample maybe brought into contact with an array of reagents such that the reagentsare printed onto the sample at known positions.

When all steps of the analysis protocol are completed on a sample in asample holder, application of the protocol may result in one or moreoptically assessable results. For example, a color of a region of thesample that was brought into contact with a dot of reagent at a knownposition on the matrix droplet extruder may be altered, or fluorescenceof the sample may be altered. In some cases, electrical properties ofthe sample, e.g., resistance or conductivity, dielectric constant, oranother electrical property of the sample, may be altered.

It may be noted that, in this manner, the composition of each reagentdot is known by its position. Therefore, it is not necessary that eachtype of reagent be distinguished from the others by a distinctlydetectable effect (e.g., by different detectable colors or electricalproperties), as would be necessary in a system where the reagents arenot spatially separated from one another and/or formed in apre-determined scheme possibly determined by the a pre-defined array ofperforations that open out of the matrix droplet extruder at the dropletmatrix extrusion surface.

After completion of all steps of an analysis protocol on a sample in asample holder, the transport mechanism may bring that sample holder to astation that includes an analysis module. For example, the analysismodule may include an imaging device (e.g., camera), illuminationsources, and appropriate optics for detecting any optical effects, e.g.,coloring or fluorescence, of a processing protocol on the sample in thesample holder.

An image of the sample may be acquired (e.g., after removal of thesample slide from the sample holder to a viewing area of the analysismodule), and the image, or a result of analysis of the image, may bedisplayed or otherwise outputted or communicated to a user. In somecases, an image or result may be stored or transmitted to another devicefor later review, analysis, or comparison. The analysis module mayinclude electronics components for measuring electrical properties ofthe processed sample.

After imaging or analysis by the analysis module, an ejection mechanismmay remove the sample holder from the transport mechanism. For example,the ejection mechanism may be configured to disconnect the sample holderfrom the transport mechanism. An inclined slide or other structure maybe configured such that an ejected sample holder may be removed from thetransport mechanism by a distance that is sufficient to prevent theejected sample holder from interfering with function of the transportmechanism or of other components of the diagnostic system.

The transport mechanism may cooperate with a loading mechanism forloading sample holders onto the transport mechanism. In some cases, theloading mechanism may be configured to hold one or more sample holdersuntil a vacancy is available on the transport mechanism. For example,the loading mechanism may include a rotatable turntable, or anothermechanical structure that may hold one or more sample holders (e.g.,capable of holding three sample holders, or another number of sampleholders) that is configured to successively place sample holders ontovacant positions on the transport mechanism as such vacant positionsbecome available.

In some cases, a reagent dot printing system may operate as a standalonesystem, and not as part of a POC diagnostic device. For example, such areagent dot printing system may include a docking station and associatedcontroller and one or more pneumatic sources (e.g., pressure actuatorsor sources). A matrix droplet extruder may be connected to the dockingstation of the reagent dot printing system. The reagent dot printingsystem may be configured to operate a liquid management chip of thematrix droplet extruder to controllably extrude an array of dots onto adroplet matrix extrusion surface of the matrix droplet extruder, wherethe matrix extrusion surface faces outward from the matrix dropletextruder. In some embodiments of the present invention the matrixdroplet extruder may be configured to controllably extrude more than onearrays (e.g., two or more arrays) of dots onto a droplet matrixextrusion surface of the matrix droplet extruder, where the matrixextrusion surface faces outward from the matrix droplet extruder.

The reagent dot printing system may be operated to print reagent dots onslides prior to their incorporation into sample holders. For example,the reagent dot printing system may be operated by a central laboratory,or by a producer of slides for a standalone sample analysis system. Thereagent dot printing system may include a mechanical system forautomatically placing slides against the droplet matrix extrusionsurface or may enable manual manipulation of the slide to the dropletmatrix extrusion surface.

A slide onto which an array of reagent dots has been printed may bestored under conditions that preserve the reagent properties of the dotsuntil used. For example, storage conditions may include one or more ofcontrolled temperature, controlled humidity, limited exposure to light,or other controlled conditions.

Prior to use, the slide may be inserted into a sample holder. Such asample holder with a preprinted slide may be utilized in a standalonesample analysis system. For example, the sample analysis system mayinclude a sample holder device that is configured to hold one or moresample holders. The sample holder device may include a controller andone or more pneumatic sources (e.g., suction pumps).

The controller may cause the flow of one or more samples or other liquidsubstances in order to filter a sample, to spread a sample on the slide,to expose the sample to one or more liquid substances (e.g., a wetprocess), or otherwise cause liquid flow within the sample holder. Thesample holder device may be configured to control liquid flow in asingle sample holder at any given time or may be configured toconcurrently control liquid flow in two or more sample holders.

For example, a sample holder device may be smaller, simpler, and lessexpensive than a POC diagnostic system. In this case, a physician, fieldworker, or other user may use such a sample holder device, together withsample holders with slides that are preprinted using a reagent dotprinting system, in a simple and efficient manner (albeit at a lowerthroughput rate), at a location that is remote from a clinic or otherinstitution with a POC diagnostic system.

FIG. 1 schematically illustrates a multiple sample analysis diagnosticdevice, in accordance with a possible embodiment of the presentinvention.

Multiple sample analysis device 10 may be configured to concurrentlyanalyze a plurality of samples. Each sample may be contained withinand/or fitted to a sample holder 18. Components of multiple sampleanalysis device 10 may be enclosed within housing 11. Multiple sampleanalysis device 10 may be utilized as a POC device.

A plurality of sample holders 18 may be inserted or fitted successivelyinto multiple sample analysis device 10 via sample holder opening 16.When analysis of a sample in a sample holder 18 is complete, that sampleholder 18 may be ejected from multiple sample analysis device 10 via anejection opening 20.

Prior to analysis of samples in sample holders 18, one or more (e.g.,one or two) matrix droplet extruders 14 may be inserted or fitted intomultiple sample analysis device 10. For example, one or more matrixdroplet extruder openings 12 may be provided to enable insertion orcoupling of a matrix droplet extruder 14. Alternatively or in addition,a cover of housing 11 may be openable to enable insertion or coupling ofone or more matrix droplet extruders 14.

A matrix droplet extruder 14 may be operated to produce an array ofdroplets of a reagent at or upon an outwardly facing surface of extruder14 that may be brought into contact with a sample that has been loadedinto a sample holder 18. For example, a single extruded droplet may havea volume between 100 pl to 2 nl, a diameter between 20 μm to 500 μm, anda spacing between dots between 100 μm to 300 μm (e.g., 200 μm betweendots of a single type of reagent, and 300 μm between differentreagents), or other values.

Multiple sample analysis device 10 may include one or more user controls22 that may be operable to control operation of multiple sample analysisdevice 10. Multiple sample analysis device 10 may include one or moreoutput devices 24 to communicate a status (e.g., number of sampleholders 18 currently inserted, whether or not another sample holder 18may be inserted, error message, and/or other status), analysis results,and/or other information to a user of the multiple sample analysisdevice 10. For example, an output device may include one or more displayscreens, indicator lights, speakers and/or other audible signalgenerators, and/or other output devices.

Multiple sample analysis device 10 may be configured to communicate viaa wired or wireless connection with an external device 28. For example,external device 28 may include a computer, terminal, or other devicesthat may enable input of instructions or data (e.g., patient or otheridentifying data associated with a sample in a sample holder 18,analysis or testing procedure or protocol to be performed on a sample ina particular sample holder 18, or other instructions), or output of dataor messages (e.g., results of analysis or testing).

Electrical power for operation of multiple sample analysis device 10 maybe provided via power connector 26. Typically, electrical power may beprovided by a direct current (DC) power source. A DC power source mayinclude an adapter that is connected to an alternating current powersource (e.g., power mains), a battery or battery pack, solar cell, orother source of DC power (e.g., with a voltage of 24 V, or anothervoltage).

FIG. 2 is a block diagram of the multiple sample analysis device shownin FIG. 1.

Transport and analysis system 40 may be configured to transport a sampleholder 18 to a plurality of predetermined stations within transport andanalysis system 40. At each station, one or more steps of an analysisprocess may be performed. After completion of the analysis process,matrix droplet extruder 14 may be ejected from multiple sample analysisdevice 10.

Operation of transport and analysis system 40 may be controlled bycontroller 36. Controller 36 may be configured to operate one or morecomponents of transport and analysis system 40 in accordance withinstructions that are generated by processor 34. Processor 34 maycommunicate with other units or devices via input/output (I/O) unit 32.For example, processor 34 may, via I/O unit 34, with one or more of auser control 22, an output device 24, and an external device 28.

Transport and analysis system 40 may include transport mechanism 70,analysis module 54, and mechanical system 50, each of which may becontrolled by controller 36.

Electrical power for operating different components of transport andanalysis system 40 and multiple sample analysis device 10 may beprovided via DC/DC converter circuitry 38. DC/DC converter circuitry 38may convert an input voltage from power connector 26 to an operatingvoltage appropriate for each component of transport and analysis system40 or of multiple sample analysis device 10.

FIG. 3A schematically illustrates an example of a transport and analysissystem of a multiple sample analysis device generally similar inprinciple to that shown in FIG. 2. FIG. 3B is a schematic exploded viewof the transport and analysis system shown in FIG. 3A. FIG. 4 is aschematic top view of the transport and analysis system shown in FIG.3A. FIG. 5 schematically illustrates a carousel of the transport andanalysis system shown in FIG. 3A.

Components of transport and analysis system 40 of multiple sampleanalysis device 10 may be mounted in this optional example to systembase 41. Mounting to system base 41 may enable components of transportand analysis system 40 to be positioned at predetermined positions andorientations relative to one another. Transport and analysis system 40may be connected to controller 36, which in turn may be configured tocontrol operation of components of transport and analysis system 40.

In the example shown, a transport mechanism 70 includes carousel 42.Carousel motor 74 may be operated to rotate carousel 42 about its axis.Carousel 42 may be configured to transport a plurality of sample holders18, up to eight in the example shown, at eight positions arranged aboutthe perimeter of carousel 42. Each sample holder 18 may be held bysample holder holding structure 19. Successive rotations of carousel 42may bring each sample holder 18 to each of an equal number of stations(e.g., eight, in the example shown) that are distributed about theperimeter of carousel 42. Carousel 42 may be operated such that eachsample holder 18 remains at each station for a predetermined period oftime (e.g., one minute, or another period of time). Thus, in the exampleshown, a rotation of carousel 42 through a rotation angle of 45° maybring each sample holder 18 from one of the stations to the nextstation.

Loading mechanism 47 may be configured to load one or more sampleholders 18 onto carousel 42. Loading mechanism 47 may include one ormore sample holder slots 48. In the example shown, loading mechanism 47includes three sample holder slots 48. For example, a sample holder 18that may be inserted or fitted into, or coupled to, sample holderopening 16 (as depicted in FIG. 1 or 5) may be placed into a sampleholder slot 48 that may be aligned or urged into correspondence withsample holder insertion opening 16.

When loading mechanism 47 includes one or more vacant sample holderslots 48, loading mechanism 47 may be rotated to align one of the sampleholder slots 48 with sample holder opening 16, in order to enableinsertion or coupling of another sample holder 18 into sample holderopening 16. When a sample holder 18 is removed from carousel 42 and thevacant sample holder holding structure 19 is rotated to the station thatincludes loading mechanism 47, a sample holder 18 may be loaded from oneof the sample holder slots 48 into sample holder holding structure 19 oncarousel 42. Thus, loading mechanism 47 may function as a buffermechanism to enable efficient loading of sample holders 18 into multiplesample analysis device 10.

A sample holder 18 may be loaded into sample holder holding structure 19on holder platform 45 to connect to a holder platform pneumaticconnector set 43. The number of sets of holder platform pneumaticconnectors 43 on holder platform 45 may, in some examples, define amaximal number of sample holders 18 (eight, in the example shown) thatmay be loaded at one time onto carousel 42.

When a sample holder 18 is loaded onto carousel 42, one or moreidentification markings on sample holder 18 may be read byidentification reader 62. For example, identification reader 62 mayinclude a barcode scanner, and RFID reader, or other sensor or devicethat may be capable of reading an identification marking on sampleholder 18. Processor 34 may be configured to associate the identifiedsample holder 18 with a source of the sample (e.g., patient or othersource) in sample holder 18, and with a possible analysis procedure thatmay be performed on or to the sample.

Each sample holder 18 may be configured to hold a sample of a liquid foranalysis. Each sample holder may include a group of pneumatic connectors(e.g., nine, or another number) that may be configured at each sampleholder position on carousel 42 to connect to (e.g., mate with) apneumatic connector set 43 possibly including an equal number ofpneumatic connectors as in the group of connectors on the holder.

Carousel 42 may include one or more pneumatic actuator systems 44 (e.g.,one pneumatic actuator system 44 to operate up to two sample holders 18,in the example shown). Each pneumatic actuator system 44 may include avacuum pump and an arrangement of conduits and valves. The number ofvalves in one possible example may be equal to the number of sampleholder positions on carousel 42, multiplied by the number of pneumaticconnectors at each sample holder position (e.g., 72 for an arrangementof eight sample holder positions and nine pneumatic connectors).

The valves may be selectively operated, e.g., by controller 36, to causea flow of one or more liquids (e.g., of a sample liquid or a liquid froma liquid reservoir of sample holder 18) through sample holder 18.Conduits that connect a pneumatic actuator system 44 with holderplatform pneumatic connectors 43 may be incorporated into holderplatform 43.

Transport and analysis system 40 may include one or more (two, in theexample shown) docking stations 56. A matrix droplet extruder 14 may beconnected to one or more of docking stations 56. A docking station 56may include a set of interfaces to a matrix droplet extruder 14 that isconnected to that docking station 56. The interfaces may include a setof pneumatic connectors for connection of a microfluidic channelingassembly within matrix droplet extruder 14 to pneumatic actuatorassembly 58.

Matrix droplet extruder 14 may include a perforated droplet matrixextrusion layer 57 (see FIG. 3B) with an outer droplet matrix extrusionsurface 571 that faces outwardly from extruder 14. Typically, dropletmatrix extrusion layer 57 may be formed as a flat slab of stainlesssteel or of another material that may be substantially nonabsorbent andcleanable. Each matrix droplet extruder 14 may be configured to extrudea matrix of reagent droplets onto droplet matrix extrusion surface 571of droplet matrix extrusion layer 57 via an array of perforations.

Matrix droplet extruder 14 may be configured to hold a plurality ofreagent containers that can be separately insertable into matrix dropletextruder 14 (e.g., in a facility for preparation of matrix dropletextruders 14, or by a user of transport and analysis system 40). Aparticular reagent may be conducted from one of the reagent containers(or reagent reservoirs into which contents of the reagent containerswere emptied) to a particular perforation on droplet matrix extrusionsurface 571 by the microfluidic channeling assembly. A configuration ofthe microfluidic channeling assembly may, in turn, be controlled bycontroller 36 via docking station 56, e.g., in accordance with a currentstep of a particular analysis that is to be performed on a sample in aparticular sample holder 18.

For example, transport and analysis system 40 may include pressureactuator 60 that is configured to generate pneumatic pressure foreffecting and controlling (via the microfluidic channeling assembly)flow of reagents to droplet matrix extrusion surface 571. For example,pressure actuator 60 may include one or more pressure pumps, and one ormore pressure regulation valves or structure.

Mechanical system 50 may be configured to manipulate sample holders 18,and other manipulable components of transport and analysis system 40.For example, mechanical system 50 may include one or more mechanicalarms 51 that may grasp and manipulate sample holders 18. Components ofmechanical system 50 may be pneumatically actuated. For example,mechanical system 50 may include a pressure pump 66. A plurality ofinternal valves may be operated to control movement of mechanical arms51 or of other components of mechanical system 50.

Rotation of carousel 42 may bring a sample holder 18 to a station atwhich a matrix droplet extruder 14 has been placed. If a sample of thatsample holder 18 is to be placed in contact with a droplet matrixextrusion surface 571 of matrix droplet extruder 14, controller 36 mayoperate a mechanical arm 51 to lift a sample slide of that sample holder18 out of sample holder 18 and to press the sample slide against dropletmatrix extrusion surface 571. For example, grasping structure 72 ofmechanical arm 51 may be configured to grasp and manipulate the slide,e.g., using structure 79 of sample slide 80 (FIG. 6A).

Typically, reagents may be extruded via perforations in droplet matrixextrusion surface 571 when the slide of sample holder 18 is in contactwith droplet matrix extrusion surface 571. In this manner, evaporationof the reagent droplets prior to contact with the sample may beprevented.

After contact of the sample with droplet matrix extrusion surface 571,droplet matrix extrusion surface 571 may be cleaned to remove any tracesof the sample prior to contact of another sample with droplet matrixextrusion surface 571. For example, mechanical arm 51 may includerinsing chamber 68. Rinsing chamber 68 may include a shallow (e.g.,having a depth of about 100 μm) indentation whose rim includes a gasket(e.g., an O-ring or other gasket).

After removal of the slide from droplet matrix extrusion surface 571,mechanical arm 51 may press rinsing chamber 68 against droplet matrixextrusion surface 571. For example, the gasket of rinsing chamber 68 mayform a seal between rinsing chamber 68 and droplet matrix extrusionsurface 571. Matrix droplet extruder 14 may then be operated to fillrinsing chamber 68 with a rinsing liquid via a rinse inlet opening indroplet matrix extrusion surface 571. After rinsing chamber 68 isfilled, matrix droplet extruder 14 may be operated to remove the rinsingliquid from rinsing chamber 68, e.g., via a rinse outlet opening indroplet matrix extrusion surface 571.

Alternatively or in addition to rinsing chamber 68, mechanical system 50or a mechanical arm 51 may be provided with one or more brushes, pads,wipers, or other structure to facilitate cleaning of droplet matrixextrusion surface 571.

Typically, after application of all steps of an analysis protocol to asample in a sample holder 18, that sample holder 18 may be rotated to astation that includes analysis module 54. For example, analysis module54 may include one or more imaging devices, light sensors, e.g, camera54 a, illumination sources, e.g. light source 54 b, or other componentsthat enable optical evaluation of a sample in sample holder 18, e.g.,after contact with an array of reagents at a droplet matrix extrusionsurface 571. Analysis module 54 may include one or more electricalcontacts, voltage or current sources, voltage or current meters,electronic circuitry, integrated circuits, or other components forelectronic evaluation of a sample in sample holder 18, e.g., aftercontact with an array of reagents at a droplet matrix extrusion surface571.

For example, when a sample holder 18 is positioned at a station thatincludes analysis module 54, mechanical arm 51 may be operated to liftthe sample slide out of sample holder 18 and place it in a viewingposition relative to analysis module 54. For example, analysis module 54may include a lightproof chamber which may enable imaging of lightreflected by or fluoresced by a sample without interference with ambientlight. An acquired image of the slide, or other data, may becommunicated to processor 34, to an external device 28, to both, or toanother destination.

Typically, after its sample is examined by analysis module 54, a sampleholder 18 may be removed from carousel 42. For example, an ejectionmechanism may remove sample holder 18 from carousel 42, such that sampleholder 18 exits multiple sample analysis device 10 via ejection opening20. For example, the ejection mechanism may include, or may cooperatewith, an inclined surface on which a sample holder 18 that is removedfrom carousel 42 may slide away from carousel 42 and out throughejection opening 20. Typically, an ejected sample holder 18 may bediscarded. In some cases, the ejected sample holder 18 may be saved forfurther analysis.

FIG. 6A schematically illustrates a sample holder of the multiple sampleanalysis device shown in FIG. 1. FIG. 6B schematically illustrates achamber for applying suction to a slide in the sample holder shown inFIG. 6A.

Components of sample holder 18 may be enclosed in holder casing 76.Holder casing 76 may be configured to connect to holder platform 45 ofcarousel 42. Pneumatic connectors 78 of sample holder 18 may connect toholder platform pneumatic connectors 43 on holder platform 45, and thusto pneumatic actuator system 44.

During application of some steps of an analysis process to a sample insample holder 18, pneumatic actuator system 44 may apply suction to oneor more pneumatic connectors 78, while one or more other pneumaticconnectors 78 may be connected to atmospheric pressure via a breathervent. Pneumatic connectors 78 may be connected to various structureswithin sample holder 18 via conduit system 88. Selective application ofsuction and/or atmospheric pressure to pneumatic connectors 78 may causeone or more liquid substances to flow between various components ofsample holder 18.

A sample may be introduced into sample holder 18 via one or more (e.g.,two or another number of) openings into sample channels 82. In theexample shown, sample channels 82 may be in the form of capillary tubes.For example, a sample chamber 82 in the form of a capillary tube may besuitable for a sample that includes blood. For other types of samples,other forms of sample channels may be provided. For example, other typesof sample channels or holders may include an absorbent or adhesivematerial strip or mass, an opening directly into a widened chamber, oranother type of channel or opening.

A sample may be introduced into a sample channel 82 after removal ofholder cover 87. For example, a sample may include a drop of blood froma pricked finger (typically a third or fourth drop after previousdroplets have been wiped away, e.g., in order to avoid contamination ofthe samples), or another liquid of biological or other origin. Once asample has been loaded into sample channels 82, holder cover 87 may bereplaced. Typically, after (or, in some cases, prior to) loading asample into sample channels 82, a barcode, RFID tag, or other identifierof sample holder 18 may be scanned and associated with an origin of thesample (e.g., a particular patient), and an analysis procedure that isto be performed on the sample by multiple sample analysis device 10.

In some cases, e.g., when the sample is a blood sample, separation maybe required. For example, blood analysis may require separating bloodcells out of the blood in sample channels 82 to yield blood plasma. Forexample, suction may be applied to sample channels 82 by pneumaticactuator system 44 via pneumatic connectors 78, plasma chamber 83, andblood separator 89 to draw blood from sample channels 82 into plasmachamber 83 via blood separator 89. For example, blood separator 89 mayinclude a filter, a microchannel, or other technique.

Structure in holder cover 87 may function as a breather vent to enableflow of the blood via blood separator 89 into plasma chamber 83. Theapplied suction (negative pressure) may be sufficiently weak so as toavoid hemolysis (rupture of blood cells). When the sample includes aliquid or solution that does not include biological cells or other typesof suspended particles that may be removed by blood separator 89, thesample may be drawn through blood separator 89 with substantially noeffect on the sample. In some cases, a sample holder 18 may be designedfor such other types of samples, e.g., without a blood separator 89.

Sample holder 18 may include sample slide 80. A sample may be applied tosubstrate 81 of sample slide 80. For example, substrate 81 may include amembrane that is composed of, or may include, nitrocellulose,fiberglass, nano-mesh, plastic, glass, or another suitable material forabsorbing a sample and other substances that may be applied to substrate81. When substrate 81 is absorbent, since the sample, or an appliedreagent, may be absorbed into substrate 81 orthogonally to its surface,the concentration of the absorbed sample or reagent per unit area may begreater than would be possible for a nonabsorbent surface. The membranemay be laid onto a web of hydrophilic/hydrophobic mesh, where thehydrophobic side of the mesh faces the membrane and the hydrophilic sideof the mesh faces away from the membrane. Wax, or other hydrophobicmaterial may be used to coat the upper side of the membrange.

A substance to be applied to substrate 81, e.g., a sample in samplechannels 82, plasma chamber 83, or a liquid substance that is held inone or more of liquid containers 84, may be drawn into sealed space 861between a section of holder casing 76 that is bounded by sealing gasket86, and a section of sample slide 80 that includes substrate 81. Thedimensions of sealed space 861 may be designed to encourage laminar flowof the sample or other liquids (e.g., to facilitate even application ofthe liquid to substrate 81.

In some cases, suction may be applied to sealed space 861 while a sourceof applied substance is opened to a breather vent, both via pneumaticconnectors 78. Once sealed space 861 has been filled by the substance,suction may be applied via chamber 77 (see FIG. 6B) to the side ofsubstrate 81 that is opposite the side that is exposed to sealed space861, while sealed space 861 is opened to a breather vent via a pneumaticconnector 78. Thus, the substance that fills sealed space 861 may bedrawn into substrate 81, thus expediting absorption of the substanceinto substrate 81.

In some cases, as illustrated schematically in FIG. 7E, the surface ofsubstrate 81 may be divided into two or more substrate sections 81′. Forexample, each substrate section 81′ may be provided with a separatesealing gasket. Alternatively or in addition, substrate 81 itself mayinclude one or more dividers 75. For example, divider 75 may be made ofa material that impedes or prevents spreading of a liquid from onesubstrate section 81′ to another. For example, different wet processsteps may be applied to two or more different substrate sections 81′. Insome cases, one or more substrate sections 81′ may function as areference or control section, e.g., to which one or more process stepsare not applied, or to which no sample is applied.

Various liquid substances may be held within sample holder 18.Typically, sample holder 18 may include between four and seven liquidcontainers 84. The number of pneumatic connectors 78 may be selected soas to enable transfer of liquids from liquid containers 84 to substrate81, with additional pneumatic connectors 78 connected to breather vents.In a non-binding example, a sample holder 18 with seven liquidcontainers 84 may include nine pneumatic connectors 78. Similarly, asample holder 18 with four liquid containers 84 may include sixpneumatic connectors 78.

A particular sample holder 18 may be configured for application of aparticular analysis procedure or of family of similar analysisprocedures, e.g., to a particular type of sample. For example, some orall sample holders 18 may be provided (e.g., by a manufacturer or userof sample holder 18) with a selection of liquid substances that are eachsealed in a blister pack 85 (to be understood as including any type ofsealed package that is insertable into a sample holder 18). Sealing of aliquid substance in a blister pack 85 may enable long-term storage ofthe liquid substance, e.g., for months or years.

Sample holder holding structure 19 of carousel 42, or a station alongthe perimeter of carousel 42, may include one or more pins orprojections (e.g., similar to projections 61 of docking station 56, asdescribed below) that are configured to be pressed against (e.g., bymechanical system 50) and puncture a blister pack 85 and emptying itscontents into a liquid container 84 for application to substrate 81(“wet process”) during analysis of the sample in sample holder 18.

Sample slide 80 may be removed from holder casing 76, e.g., by amechanical arm 51 or by another component of mechanical system 50. Forexample, substrate 81 of sample slide 80 may be placed against dropletmatrix extrusion surface 571 of matrix droplet extruder 14 or may beplaced into analysis module 54. Sample slide 80 (see FIGS. 6A and 6B)may include registration structure 79 to facilitate precise registrationwith droplet matrix extrusion surface 571 or analysis module 54.

For example, registration structure 79 may include mechanical structure(e.g., holes, indentations, pins, projections, or other mechanicalstructure), optical structure (e.g., reflectors, colored lines, bars,dots, patterns, or other optical structure), electromagnetic structure,or other structure to enable precise registration of sample slide 80with cooperating structure, e.g., on droplet matrix extrusion surface571, matrix droplet extruder 14, analysis module 54, or elsewhere.

FIG. 7A schematically illustrates a matrix droplet extruder insertedinto a docking station that may be used in the transport and analysissystem shown in FIG. 3A. FIG. 7B is a schematic opposing view of thedocking station shown in FIG. 7A. FIG. 7C schematically illustrates arear side of the matrix droplet extruder partially seen in FIGS. 7A and7B. FIG. 7D is a schematic cross section of the matrix droplet extrudershown in FIG. 7C.

Docking station 56 in this example includes insertion space 67 intowhich a matrix droplet extruder 14 may be inserted. For example, matrixdroplet extruder 14 may be handled (e.g., without contaminating dropletmatrix extrusion surface 571) using pack handle 15.

Casing 91 (see FIG. 7C) of matrix droplet extruder 14 may include aplurality of docking pneumatic connectors 92 that may be connectable todocking station connectors 65 (see FIG. 7B) within insertion space 67 ofdocking station 56. Internal structure of matrix droplet extruder 14,e.g., within casing 91, may be controlled by pneumatic actuator assembly58 via docking pneumatic connectors 92.

Matrix droplet extruder 14 may include a plurality of reagent containers93 (see FIG. 7D), each for holding a liquid reagent or other liquids(e.g., cleaning liquids, hydraulic liquids, or other liquids, all ofwhich are referred to herein as reagents for convenience and clarity).Reagent liquids may be provided in reagent canisters 94. For example, areagent canister 94 may include a blister pack or other sealed containerthat may be emptied into a reagent container 93.

Prior to attachment to docking station 56, a reagent canister 94 may beplaced in each or some or all of reagent containers 93, e.g., by amanufacturer, distributor, or user of matrix droplet extruder 14. Aselection of reagent canisters 94 (e.g., each containing a particularliquid reagent) for insertion into reagent containers 93 may beconfigured for a particular facility or type of facility (e.g., clinic,mobile facility, hospital, or other type of facility) or for aparticular purpose (e.g., blood test or other clinical tests, chemicalevaluation of substances, or other purpose), or otherwise.

A liquid reagent may be stored in a reagent canister 94 for a longperiod (e.g., months or years). (It should be understood that referenceherein to reagent containers, canisters, conduits, or other reagentcomponents or structure should be understood as referring to componentsor structure for holding, or directing the flow of, any liquid thatutilized during operation of matrix droplet extruder 14.)

After insertion of matrix droplet extruder 14 into insertion space 67,activation plate 59 of docking station 56 may be operated to preparematrix droplet extruder 14 for operation, e.g., at the beginning of awork period (e.g., a work day), or otherwise after insertion of matrixdroplet extruder 14 and prior to operation of matrix droplet extruder14. A mechanism may push activation plate 59 forward toward the rearside (e.g., the side opposite of that which includes droplet matrixextrusion surface 571) of matrix droplet extruder 14. In the exampleshown, activation plate 59 may be pushed forward by application ofpneumatic pressure to plate movement pneumatic connectors 63.

Activation plate 59 may include a plurality of projections 61, e.g.,cylindrical pins or other projections. Pushing activation plate 59 tomatrix droplet extruder 14 may insert projection 61 into a reagentcanister 94. Insertion of projection 61 may rupture a part of eachreagent canister 94 or otherwise force the contents of that reagentcanister 94 into the reagent container 93 into which that reagentcanister 94 was inserted.

Typically, a liquid reagent may be stored in a reagent container 93 forthe duration of a typical work period (e.g., one day). In some cases,multiple sample analysis device 10 may be provided with refrigeration,either internal (e.g., refrigeration structure enclosed within housing11) or external (e.g., a refrigerated case into which multiple sampleanalysis device 10 or matrix droplet extruder 14 may be inserted), toextend a usable lifetime of the liquid reagents (e.g., for use in afacility with a small daily throughput of samples).

In some cases, docking station 56 may be configured for separateinsertion of subsets of projections 61 into a subset of reagentcanisters 94. For example, docking station 56 may include a plurality ofseparately operable activation plates 59 or may include a mechanism forselectively extending, retracting, or otherwise operating individualprojections 61 or groups of projections 61.

In addition to emptying the contents of reagent canisters 94, pushingactivation plate 59 into matrix droplet extruder 14 may also cause eachof docking station connectors 65 to connect to a corresponding dockingpneumatic connector 92 (see FIG. 7C) on matrix droplet extruder 14.Thus, pneumatic actuator assembly 58 may be operated to providepneumatic pressure to matrix droplet extruder 14 via docking pneumaticconnectors 92.

When matrix droplet extruder 14 is to print a pattern of reagent dotsvia droplet matrix extrusion surface 571, pneumatic actuator assembly 58may be operated to force a liquid reagent out of a reagent container 93,via reagent channels of channel arrangement 96 (see FIG. 7D), to ortowards liquid management chip 98. Liquid management chip 98 may becontrollable via a plurality of pneumatic control channels 97 of channelarrangement 96 that connect each of one or more docking pneumaticconnectors 92 to one or more pneumatically operable gates 95 of liquidmanagement chip 98.

Liquid management chip 98 may include an internal arrangement of controlconduits and dispensing channels 99 so as to control and direct a flowof liquid reagents from reagent containers 93, via channel arrangement96, to specific perforations 69 of droplet matrix extrusion surface 571.For example, channel arrangement 96 and dispensing channels 99 may bearranged so as to connect a particular reagent container 93 to one or,more typically, to a plurality of specific perforations 69 of dropletmatrix extrusion surface 571. The reagent may be dispensed whenpneumatic pressure is applied to the reagent, e.g., at reagent container93. Closing a pneumatically operable gate 95 may prevent extrusionthrough a particular perforation 69.

In some cases, two or more reagent containers 93 may be connected to asingle perforation 69. For example, one of the two or more reagentcontainers 93 may be filled initially, while another is filled when thefirst is emptied or when a predetermined period of time has passed sincebeing opened. In some cases, a cleaning fluid may be extruded viaperforation 69 between successive extrusions of reagent from thedifferent reagent containers 93.

In some cases, extrusion of two or more different reagents via a singleperforation 69 may enable formation of a droplet that includes a mixtureor other combination of both reagents. For example, a perforation 69 maybe elongated to enable connection to two or more dispensing channels 99.As another example, droplet matrix extrusion surface 571 may includegrooves or other structure that enables or facilitates mixing (e.g., viaone or more channeling effects) between different reagents that areextruded by two or more separate but neighboring perforations 69.

Liquid management chip 98 may be controlled by pneumatic actuatorassembly 58 (see indicated in FIGS. 3 and 4) to direct a particularliquid reagent from a selected reagent container 93 to one or moreselected perforations 69 on droplet matrix extrusion surface 571. Theliquid reagent may be extruded via that perforation 69, e.g., to printthat reagent onto a sample on substrate 81 of a sample slide 80.

FIG. 7E schematically illustrates an example of a slide onto whichreagent dots were extruded.

In the example shown, reagent dots 71 have been printed on substrate 81.Although, in the example shown, substrate 81 is divided by dividers 75into multiple substrate sections 81′, substrate 81 may be a singleundivided surface (e.g., membrane). In the example shown, reagent dots71 are organized into dot rows 73, where the separation betweenneighboring dot rows 73 is here optionally greater than the separationbetween adjacent reagent dots 71 in a single dot row 73. For example,each dot row 73 may include reagent dots 71 of a single reagent (e.g.,where any mixing between adjacent reagent dots 71 is less likely toaffect results), while different dot rows 73 may be of differentreagents where mixing is to be avoided.

Alternatively or in addition to organization of reagent dots 71 intoseparate dot rows 73, groups of reagent dots 71 may be organized intoseparated columns, clusters, or otherwise, or without separationsbetween groups. In other examples, parts of substrate 81 may be coatedor infused with a liquid repellent (e.g., using a solid ink printer) tolimit absorption to predetermined regions of substrate 81 (e.g., tocreate separate assay regions on substrate 81).

FIG. 8 schematically illustrates transport of sample holders to stationsof the transport and analysis system shown in FIG. 3.

Stations A-H are distributed about the perimeter of carousel 42.Transport mechanism 70 may be configured to rotate in the directionshown by the arrow to transport each loaded sample holder 18 from one ofstations A-H to a succeeding station B-A, respectively. After a sampleholder 18 is transported to a succeeding station, carousel 42 remainsstationary (e.g., does not rotate) for a predetermined period of time.

FIG. 9 is a flowchart depicting a method of operation of a multiplesample analysis device, in accordance with an embodiment of the presentinvention.

It should be understood with respect to any flowchart referenced herein,that the division of the illustrated method into discrete operationsrepresented by blocks of the flowchart has been selected for convenienceand clarity only. Alternative division of the illustrated method intodiscrete operations is possible with equivalent results. Suchalternative division of the illustrated method into discrete operationsshould be understood as representing other embodiments of theillustrated method.

Similarly, it should be understood that, unless indicated otherwise, theillustrated order of execution of the operations represented by blocksof any flowchart referenced herein has been selected for convenience andclarity only. Operations of the illustrated method may be executed in analternative order, or concurrently, with equivalent results. Suchreordering of operations of the illustrated method should be understoodas representing other embodiments of the illustrated method.

POC sample analysis method 100 may be executed by a processor, such asprocessor 36 of multiple sample analysis device 10, or by anotherprocessor in communication with processor 36 or with multiple sampleanalysis device 10.

A sample holder 18 may be received (block 110). For example, sampleholder 18 may have been loaded into multiple sample analysis device 10via sample holder insertion opening 16, and may have been loaded ontotransport mechanism 70, e.g., carousel 42. In the example shown in FIG.8, a sample holder 18 is loaded onto carousel 42 at station D, e.g., vialoading mechanism 47.

Each sample holder 18 is transported by transport mechanism 70 to asucceeding station (block 120), where the sample holder 18 remains for apredetermined period of time (e.g., one minute, or another appropriateperiod of time). For example, a sample holder 18 that was loaded ontocarousel 42 at station D may be transported to station E. The othersample holders 18 on carousel 42 are similarly transported to asucceeding station.

An action may be performed on a sample holder 18 that is remaining ateach of the stations for the predetermined period of time (block 130).The action that is performed may depend on a current step of an analysisprocess that is being performed on the sample held in each sample holder18.

For example, steps of an ELISA protocol may begin to be applied to asample in a sample holder 18 when that sample holder 18 is at station A.Succeeding steps of the ELISA protocol may be performed at eachsucceeding station B-G (depending on the type or ELISA protocol, e.g.,indirect or sandwich Elisa protocol). In this case, and when the sampleis a blood sample, the blood may be caused to flow from sample channels82 through blood separator 89 at each of stations E-H between loading ofsample holder 18 at station D and application of the first step of theELISA protocol at station A.

In some cases, the performed action may include waiting at the stationwithout any processing of the sample held in sample holder 18. In somecases, waiting at a station may provide an incubation step of an ELISAprotocol.

In some cases, the action may include placement of the sample intocontact with an array of reagent droplets on a droplet matrix extrusionsurface 571 of a matrix droplet extruder 14, at station B or F in theexample shown. When a sample holder 18 has been transported to station Bor F, mechanical arms 51 may lift sample slide 80 out of sample holder18 and place substrate 81 into contact with droplet matrix extrusionsurface 571. For example, when the sample includes, or is suspected ofincluding, one or more specific antigens, the array of reagent dropletsmay include one or more specific detection antibodies to which detectionmolecules (e.g., a fluorescent or other dye, such a horseradishperoxidase) have been attached. Typically, the specific contents of eachreagent droplet is duplicated, e.g., in a row or column of adjacentreagent droplets.

Following removal of slide 80 from droplet matrix extrusion surface 571,mechanical system 50 may place rinsing chamber 68 over droplet matrixextrusion surface 571. Pneumatic actuator assembly 58 may then beoperated to cause a cleaning fluid to flow from a cleaning fluidreservoir of matrix droplet extruder 14 via a cleaning fluid inlet intorinsing chamber 68. After a predetermined period of time, pneumaticactuator assembly 58 may be operated to drain the cleaning fluid fromrinsing chamber 68 via a cleaning fluid outlet into a waste fluidreservoir of matrix droplet extruder 14. Mechanical system 50 may removerinsing chamber 68 from droplet matrix extrusion surface 571.

At at least some stations, a sample may be subjected to one or more wetprocesses. For example, a substrate 81 to which an ELISA protocol isapplied may be subjected to one or more coating, washing, blocking, orother steps. In particular, a coating step may include coating with thesample, or coating with a capture antibody prior to coating with thesample.

When a sample holder 18 is transported to a station that includesanalysis module 54 (station H in the example shown), e.g., after some orall processing steps have been applied to the sample, the sample may beexamined by analysis module 54. For example, mechanical system 50 may becontrolled to lift sample slide 80 out of sample holder 18 so thatsample slide 80, or at least substrate 81, is inserted into analysismodule 54, e.g., a light tight chamber of analysis module 54.

Analysis module 54 may then be operated to acquire one or more images ofthe substrate 81, e.g., under various types of illumination, orfluorescence of substrate 81. Alternatively or in addition, analysismodule 54 may apply electronics to measure one or more electricalproperties of the locations on substrate 81. The analysis may includedetermining a correspondence of a result at each location on substrate81 with a reagent with which substrate 81 was placed into contact atthat location.

Analysis may include mapping a measured property of the substrate withina reagent dot (e.g., which may have an irregular shape). The analysismay include combining measurements at different points of a reagent dot,or different reagent dots of the same reagent, to calculate astatistically significant result. The analysis may include preparing areport that summarizes the results of the analysis.

If all actions on a sample holder 18 are complete (block 140),mechanical system 50 may be operated to remove that sample holder 18from carousel 42 and to eject it from multiple sample analysis device 10(block 150). Typically, the final action performed on a sample holder 18is examination of its sample slide 80 by analysis module 54.

If additional actions remain to be performed on a sample holder 18(block 140), that sample holder 18 is transported to the next station(block 120), and an additional action may be performed (block 130).

A matrix droplet extruder 14 may be incorporated into a standalonereagent dot printing system for preprinting reagent dots on a sampleslide 80. Similarly, a preprinted slide may be incorporated into asample holder 18 for use with a standalone sample analysis system.

FIG. 10 schematically illustrates an example of a reagent dot printingsystem.

Reagent dot printing system 200 may be utilized to print an array ofreagent dots 71 on a surface. In particular, reagent dot printing system200 may be operated to print an array of reagent dots 71 on a slide toproduce a preprinted sample slide 204 on which reagent dots 71 arepre-extruded. As used herein, pre-extruded refers to extrusion ofreagent droplets onto substrate 81 prior to introduction of the sampleonto that substrate 81.

For example, dot printing device 202 may incorporate a docking station56. Dot printing device 202 may include one or more pneumatic actuators203 and conduits for operation of a matrix droplet extruder 14 that isconnected to docking station 56. Dot printing device 202 may incorporateor may communicate with a controller 201 for controlling operation ofthe pneumatic actuators. For example, controller 201 may be configuredto control a pressure that is generated by pressure actuator 60 (notshown in FIG. 10) of pneumatic actuators 203 and that is applied to oneor more docking pneumatic connectors 92 of matrix droplet extruder 14,or time of operation of the pressure actuator 60 or a period of timeduring which the pressure is applied to one or more docking pneumaticconnectors 92.

In some cases, dot printing device 202 may be provided with useroperable controls for controlling operation of dot printing device 202.In some cases, operation of dot printing device 202 may be controlledvia an external device 28 (e.g., computer, smartphone, or other device)that communicates with dot printing device 202 via a wired or wirelessconnection.

For example, dot printing device 202 may be operated (e.g., by apharmacist, physician, or other trained user) to produce a preprintedsample slide 204. Typically, dot printing device 202 may be operated toproduce a plurality of preprinted sample slides 204 that are designedfor use by a particular user (person or institution), under particularcircumstances (e.g., for application of a particular protocol, or fordetection of a particular physiological disorder, toxin, contaminant, orother component of a liquid sample), geographic location (e.g., whereparticular conditions are prevalent), or other particular application.

Preprinted sample slides 204 may be immediately inserted into sampleholders 18 or stored separately for later insertion into sample holders18 or for other uses. Storage of preprinted sample slides 204, or ofsample holders 18 into which preprinted sample slides 204 have beeninserted, may be under controlled conditions designed to preserve theintegrity of reagent dots that are printed onto preprinted sample slide204.

FIG. 11 schematically illustrates an example of a standalone sampleprocessing system.

Sample processing system 210 is configured to perform one or moreprocess steps, and, in particular, wet process steps, on a sample thatis introduced into sample holder 18. For example, sample processingdevice 212 may include pneumatic actuators to cause liquid flow of oneor more samples or other liquids within sample holder 18 that isinserted into sample processing device 212.

When a sample holder 18 includes a preprinted sample slide 204, theprocessed sample may interact with the reagent dots on preprinted sampleslide 204 to produce an optically assessable result (e.g., color changeor fluorescence). Typically, sample processing device 212 includes ananalysis module 54 for optically or electronically examining apreprinted sample slide 204 after the processing that occurs within asample holder 18 that is inserted into sample processing device 212. Insome cases, sample processing device 212 may include a mechanical system(e.g., with some components of mechanical system 50 as described above)to remove preprinted sample slide 204 from sample holder 18. In othercases, a user may remove preprinted sample slide 204 from sample holder18 for analysis by analysis module 54.

For example, sample processing device 212 may be utilized by a privateuser (e.g., a patient) or other user who does not need to analyze alarge number of samples. Thus, a user of sample processing device 212may purchase preprinted sample slides 204 for a particular test, or fora limited number of tests, and utilize appropriate sample holders 18 toperform the appropriate wet process steps on the sample.

According to some embodiments of the present invention, a sampleanalysis device may include a sample holder and a matrix dropletextruder configured to controllably extrude two or more arrays of dotsonto a droplet matrix extrusion surface of the matrix droplet extruder,where the matrix extrusion surface faces outward from the matrix dropletextruder. The sample holder in such embodiments may include a sampleslide with two or more isolated zones, and the sample analysis devicemay be configured to manipulate the sample slide so as to performprimary printing of an array of droplets onto said two or more isolatedzones, for example, of a first antibody agent or agents, followed bysecondary printing of a second antibody agent or agents onto said two ormore isolated zones on the sample slide. According to some embodimentsof the invention various preparatory steps may be performed in betweenthe two printings, e.g., washing, blocking, and other steps.

FIG. 12A is a front view of a sample holder 1200 of the multiple sampleanalysis device, according to some embodiments of the present invention,with a sample slide inserted inside. FIG. 12B is a front view of asample holder 1200 of the multiple sample analysis device, according tosome embodiments of the present invention, with the sample slide pulledout.

Components of sample holder 1200 may be enclosed in holder casing 76 a.Holder casing 76 a may be configured to connect to holder platform 45 ofcarousel 42 or another platform, and may be movable or fixed inposition. Pneumatic connectors 78 a of sample holder 1200 may connectvia holder platform pneumatic connectors to a pneumatic actuator system.

During application of some steps of an analysis process to a sample insample holder 1200, the pneumatic actuator system may apply suction toone or more pneumatic connectors 78 a, while one or more other pneumaticconnectors 78 a may be connected to atmospheric pressure via a breathervent. Pneumatic connectors 78 a may be connected to various structureswithin sample holder 1200 via a conduit system 88 a. Selectiveapplication of suction and/or atmospheric pressure to pneumaticconnectors 78 a may cause one or more liquid substances to flow betweenvarious components of sample holder 1200.

A sample may be introduced into sample holder 1200 via one or more(e.g., two or another number of) openings into sample channel (orchannels) 82 a. Sample channel 82 a may be in the form of capillarytube, e.g., capillary tube suitable for a sample that includes blood.For other types of samples, other forms of sample channels may beprovided. For example, other types of sample channels or holders mayinclude an absorbent or adhesive material strip or mass, an openingdirectly into a widened chamber, or another type of channel or opening.

A sample may be introduced into a sample channel 82 a. For example, asample may include a drop of blood from a pricked finger (typically athird or fourth drop after previous droplets have been wiped away, e.g.,in order to avoid contamination of the samples), or another liquid ofbiological or other origin. Once a sample has been loaded into samplechannel 82 a, a holder cover 87 a may be placed on the inlet of thesample channel 82 a to seal it. Typically, after (or, in some cases,prior to) loading a sample into sample channel 82 a, a barcode, RFIDtag, or other identifier of sample holder 1200 may be scanned andassociated with an origin of the sample (e.g., identity of a particularpatient), and an analysis procedure that is to be performed on thesample by a multiple sample analysis device.

In some cases, e.g., when the sample is a blood sample, separation maybe required. For example, blood analysis may require separating bloodcells out of the blood in sample channel 82 a to yield blood plasma. Forexample, suction may be applied to sample channel 82 a by the pneumaticactuator system via pneumatic connectors 78 a, plasma chamber 83 a, andblood separator 89 a to draw blood from sample channel 82 a into plasmachamber 83 a via blood separator 89 a. For example, blood separator 89 amay include a filter, a microchannel, or another device employinganother separation technique.

Holder cover 87 a may include a breather vent to enable flow of theblood via blood separator 89 a into plasma chamber 83 a. In some otherembodiments, sample holder 1200 may be designed for such other types ofsamples.

Sample holder 1200 may include sample slide 80 a configured to beinserted into or removed from a pocket 1201 in sample holder 1200. Asample may be applied to two or more substrate patches 81 a and 81 b ofsample slide 80 a. For example, substrate patches 81 a and 81 b mayinclude, each, a membrane that is composed of, or may include,nitrocellulose, fiberglass, nano-mesh, plastic, glass, or anothersuitable material for absorbing a sample and other substances that maybe applied to substrate patches 81 a and 81 b. When substrate patches 81a and 81 b are absorbent, since the sample, or an applied reagent, maybe absorbed into substrate patches 81 a and 81 b orthogonally to theirsurface, the concentration of the absorbed sample or reagent per unitarea may be greater than would be possible for a nonabsorbent surface.

When sample slide 80 a is inside pocket 1201 in sample holder 1200,substance to be applied to substrate patches 81 a and 81 b, e.g., asample in sample channel 82 a, plasma chamber 83 a, or a liquidsubstance that is held in one or more of liquid containers 84 a, may bedrawn into sealed spaces 861 a and 861 b between a section of holdercasing 76 a that is bounded by sealing gasket 86 a, and either of thesections of sample slide 80 a that include substrate patch 81 a orsubstrate patch 81 b. The dimensions of sealed spaces 861 a and 861 bmay be designed to obtain laminar flow of the sample or other liquids(e.g., to facilitate even application of the sample or one or more kindsof the other liquid to substrate patches 81 a and/or 81 b).

In some cases, suction may be applied to sealed spaces 861 a and 861 bwhile a source of applied substance is opened to a breather vent, bothvia pneumatic connectors 78 a. Once sealed spaces 861 a and/or 861 bhave been filled by the substance, suction may be applied via chamber 77(see FIG. 6B) to the side of substrate 81 a that is opposite the sidethat is exposed to sealed spaces 861 a and 861 b, while sealed spaces861 a and 861 b are opened to a breather vent via a pneumatic connector78 a. The substance that fills sealed spaces 861 a and 861 b may bedrawn into substrate patches 81 a and 81 b respectively, thus expeditingabsorption of the substance into substrate patches 81 a and 81 b.

Various liquid substances may be held within sample holder 1200.Typically, sample holder 1200 may include a plurality of liquidcontainers 84 a. The number of pneumatic connectors 78 a may be selectedso as to enable transfer of liquids from liquid containers 84 a tosubstrate patches 81 a and 81 b, with additional pneumatic connectors 78a connected to breather vents.

A particular sample holder 1200 may be configured for application of aparticular analysis procedure or of family of similar analysisprocedures, e.g., to a particular type of sample. For example, some orall sample holders 1200 may be provided (e.g., by a manufacturer or userof sample holder 18) with a selection of liquid substances that are eachsealed in a blister pack 85 a (may include any type of sealed packagethat is insertable into a sample holder 1200).

Sample slide 80 a may include a locking arrangement that locks thesample slide 80 a in position when the sample slide 80 a is fullyinserted inside the sample holder 1200. For example lock 1220 may beincluded, which when the sample slide 80 a is fully inserted inside thesample holder 1200, lock 1220 is operated, e.g., by a mechanical arm, tosecure the sample slide 80 a in its position, when it is desired toremove the sample slide 80 a from the sample holder 1200, lock 1220 isrelease, e.g., by the mechanical arm, and the sample slide 80 a isreleased and may be removed from sample holder 1200.

FIG. 13A shows a matrix droplet extruder configured to perform dualprinting assays, according to some embodiments of the presentapplication, with a sample slide in a first print position.

FIG. 13B shows a matrix droplet extruder configured to perform dualprinting assays, according to some embodiments of the presentapplication, with a sample slide in a second print position.

Matrix droplet extruder 1300 may be used in a sample analysis systemaccording to some embodiments of the present invention.

FIG. 13C shows the back of matrix droplet extruder 1300, according tosome embodiments of the present invention.

Casing 91 a of matrix droplet extruder 1300 may include a plurality ofdocking pneumatic connectors 92 a that may be connectable to dockingstation connectors (e.g., docking station connectors 65, see FIG. 7B) ofa docking station (e.g. docking station 56). The internal structure ofmatrix droplet extruder 1300, e.g., within casing 91 a, may becontrolled by a pneumatic actuator assembly (e.g., pneumatic actuatorassembly 58 via docking pneumatic connectors 92).

Matrix droplet extruder 1300 may include a plurality of reagentcontainers (e.g. containers 93 a), each for holding a liquid reagent orother liquids (e.g., cleaning liquids, hydraulic liquids, or otherliquids, all of which are referred to herein as reagents for convenienceand clarity). Reagent liquids may be provided in reagent containers ofvarious types, e.g. canisters. For example, a reagent canister mayinclude a blister pack or other sealed container that may be emptiedinto a reagent container.

Matrix droplet extruder 1300 may include a liquid management chip withan internal arrangement of control conduits and dispensing channels soas to control and direct a flow of liquid reagents from reagentcontainers, via the channel arrangement, to specific perforations 69 aof droplet matrix extrusion surface 571 a. the channel arrangement andthe dispensing channels may be arranged so as to connect a particularreagent container to one or, more typically, to a plurality of specificperforations 69 a of droplet matrix extrusion surface 571 a. The reagentmay be dispensed when pneumatic pressure is applied to the reagent,e.g., at the reagent container. Closing a pneumatically operable gatemay prevent extrusion through a particular perforation 69 a.

The extrusion surface 571 a of matrix droplet extruder 1300 may includetwo (or more) separate printing zones 1302 a and 1302 b, each of theprinting zones comprising a plurality of perforations 69 a. The separateprinting zones may serve in an analysis process that requires more thanone printing of reagents on the sample on the sample slide.

A washer 1304 may be provided, for washing the printing zones 1302 a and1302 b on the matrix extrusion surface 571 a. The washer may bepositioned in a stand-by (idle) position (as shown in FIGS. 13A and13B), when the sample slide 80 a is placed over the printing zones. Whenwashing of the printing zones is required, the washer may be transported(e.g., by a mechanical arm, or by another transporting mechanism) to aposition over the printing zones 1302 a and 1302 b of the matrixextrusion surface 571 a, to perform washing.

Washer 1304 may include in inlet port 1306 for introducing a washingagent (e.g., cleaning fluid) onto the printing zones and for evacuatingthe washing agent from the printing zones.

When the sample slide 80 a is not over the printing zone the washer 1304may be placed over the printing zones in order to maintain a humidenclosure, bound by the gaskets 1308 a and 1308 b, over perforations 69a, so as to prevent perforation blockage caused by dried up substance.

A matrix droplet extruder such as depicted in FIGS. 13A and 13B may besuitable for use in a sample analysis system, which requires dual (ormore) printing. For example, an analysis protocol (e.g., ELISA) mayrequire introduction of a first antibody to a sample, and laterintroduction of a secondary antibody to the sample. Such protocol stepsmay include printing a first set of antibody or antibodies on the sampleslide, binding that first antibody, washing remains of the firstantibody that did not bind, and printing a second set of a secondaryantibody.

In some embodiments of the present invention a sample analysis systemmay include one or a plurality of matrix droplet extruders, such as theone depicted in FIGS. 13A and 13B. Having two or more matrix dropletextruders such as the one depicted in FIGS. 13A and 13B may facilitateperforming various analysis protocols that require combinations ofprinting options. Thus, for example, it may be possible to perform, foreach patient's sample, an analysis protocol having two or moreoverlapping printing assays, or having a plurality of different tests(e.g., upper printing zone of first matrix droplet extruder to test fora plurality of biomarkers—a biomarker test panel A, lower printing zoneof that first matrix droplet extruder to test for biomarker test panelB, upper printing zone and lower printing zone of second matrix dropletextruder to test for biomarker test panel C—requiring dual printingassays, etc.).

A sample analysis system, according to some embodiments of the presentinvention may include a matrix droplet extruder such as the one depictedin FIGS. 13A and 13B. The sample slide 80 a may be removed from thesample holder 1200, placed over the first printing zone 1302 a, whereprinting of a first array of the first antibody is performed onto thesample slide (e.g., sample membrane). The sample slide 80 a may then beremoved from the first printing position, returned into the sampleholder 1200, may undergo additional processing (e.g., washing) and thenremoved again from the sample holder 1200 and placed over the secondprinting zone 1302 b, where printing of the a second array of secondaryantibody is performed over the printed first array of the firstantibody. When placed over one of the printing zones, bores 1227 insample slide 80 a (see FIG. 12B) may be used to cooperate with matchingpins to properly align (register) the sample slide 80 a over theappropriate printing zone.

Some embodiments of the present invention may be used in performingvarious assays, e.g., direct assay, indirect assay, sandwich assay, etc.

FIG. 13D is a front view of a washer 1304 of the matrix droplet extrudershown in FIG. 13A and FIG. 13B.

Washer 1304 may include two (or more) insulated washing enclosures 1310a and 1310 b, which are separated from each other, for example, by meansof gaskets 1308 a and 1308 b insulatingly encircling the washingenclosures 1310 a and 1310 b, respectively.

Washing agent (e.g., liquid) may be introduced into the washingenclosures 1310 a and 1310 b and evacuated from these enclosures, in oneor a plurality of cycles to wash the printing zones 1302 a and 1302 b,when the washer is placed over these printing zones and operated. Thewasher may be connected to a first container for containing washingliquid, and to a second container for containing the washing agent afterit was used to wash the printing zones.

FIG. 14 is a schematic illustration of a sample analysis system 1400,according to some embodiments of the present invention.

Sample analysis system 1400 may include a housing 1420 which houses amatrix droplet extruder 1300, a transport mechanism, that may include alinear displacing mechanism 1404 for moving a sample holder 1200 on amovable docking station 1415 from an input port 1402 to a positionfacing matrix droplet extruder 1300, or for moving the sample holder1200 to docking station 1415 facing the matrix droplet extruder 1300,and a lifting mechanism 1406 for engaging with sample slide 80 a and forlifting it to at least one printing position over the extrusion surfaceof matrix droplet extruder 1300. In some embodiments lifting mechanism1406 is configured to engage with sample slide 80 a and to lift it to afirst printing position and/or to a second printing position over theextrusion surface of matrix droplet extruder 1300. After printing iscompleted the lifting mechanism and/or the displacing mechanism 1404places sample slide 80 a in front of imaging device 1416 (e.g., camera)for the imaging device to grab an image of the sample slide 80 a. Afterhe obtaining the image, the sample holder 1200 with the sample slide 80a are discarded through port 1422 into disposal container 1424, andretained in there. Controller 1410 is provided, configured to controlthe operation of the displacing mechanism 1404, the lifting mechanism1406, docking station 1415, and imaging device 1416.

The transport mechanism (e.g., displacing mechanism and or the liftingmechanism) may comprise track or tracks, motor and transmission, chain/sand/or belt/s, and/or any other known mechanism or mechanisms fordisplacing and/or for lifting and lowering objects.

A sample analysis system 1400, according to some embodiments of thepresent invention, may be used in a point of care, to perform varioustests om biological samples (e.g., blood, urine, saliva, etc.) ofpatients. Each patient may be assigned a single sample holder into whicha sample to be tested of that patient is loaded. The sample slide ofeach patient undergoes various testing stages, for example, such asdescribed hereinabove, and when the testing is completed the sampleholder with the sample slide of that patient may be discarded into thedisposal container. From time to time, or when the disposal container isfilled, it may be emptied via port 1426, or the entire disposalcontainer may be replaced with a new empty disposal container. The usedsample holders with the used sample slides may preferably be treated forbiological waste in accordance to proper medical standards.

Different embodiments are disclosed herein. Features of certainembodiments may be combined with features of other embodiments; thus,certain embodiments may be combinations of features of multipleembodiments. The foregoing description of the embodiments of theinvention has been presented for the purposes of illustration anddescription. It is not intended to be exhaustive or to limit theinvention to the precise form disclosed. It should be appreciated bypersons skilled in the art that many modifications, variations,substitutions, changes, and equivalents are possible in light of theabove teaching. It is, therefore, to be understood that the appendedclaims are intended to cover all such modifications and changes as fallwithin the true spirit of the invention.

While certain features of the invention have been illustrated anddescribed herein, many modifications, substitutions, changes, andequivalents will now occur to those of ordinary skill in the art. It is,therefore, to be understood that the appended claims are intended tocover all such modifications and changes as fall within the true spiritof the invention.

1-8. (canceled)
 9. A sample holder comprising: a casing with a pocket; a removable sample slide with one or a plurality of substrate patches configured to be inserted into or removed from the pocket; a sample channel for introducing a liquid sample into one or a plurality of sealed spaces between the sample slide and a section of the casing, so as to allow the sample liquid to be absorbed in said one or more substrate patches.
 10. The sample holder of claim 9, wherein the substrate patches comprise a membrane.
 11. The sample holder of claim 10, wherein the membrane comprises material selected from the group of materials consisting of: nitrocellulose, fiberglass, nano-mesh, plastic and glass.
 12. The sample holder of claim 9, further comprising a plurality of pneumatic connectors on the casing that are each configured to be connected to a pneumatic pressure or vacuum actuator to provide controlled pneumatic pressure or vacuum to one or more of said plurality of the pneumatic connectors to obtain laminar flow of the sample or other liquids into said one or a plurality of sealed spaces.
 13. The sample holder of claim 12, further comprising one or a plurality of liquid containers containing one or more kinds of the other liquids.
 14. The sample holder of claim 9, further comprising a lock for locking the sample slide when fully inserted inside the pocket.
 15. A sample analysis system comprising: a pneumatic actuator; one or a plurality of sample holders comprising: a casing with a pocket; liquid containers; a removable sample slide with a substrate comprising one or a plurality of substrate patches, configured to be inserted into or removed from the pocket; and a sample channel for introducing a liquid sample into one or a plurality of sealed spaces between the sample slide and a section of the casing, so as to allow the sample liquid to be absorbed in said one or more substrate patches; and a controller that is configured to control the pneumatic actuator so as to sequentially cause one or more liquids from one or more of the liquid containers to flow to or away from the sample slide in that sample holder; and an analysis module for examining the sample slide after contact of the sample with a plurality of dots of one or more reagents that are printed onto the substrate.
 16. The system of claim 15, further comprising a transport mechanism to transport each of said one or a plurality of the sample holders sequentially to a station of a plurality of stations along the transport mechanism, the controller further configured to control the transport mechanism so as to cause the transport mechanism to move said one or a plurality of sample holders such that each of the one or plurality of sample holders is sequentially moved from a current station of the plurality of stations to a next station of the plurality of stations.
 17. The system of claim 16, wherein the transport mechanism comprises a rotatable carousel, the plurality of pneumatic connectors arranged to enable connection of each of said one or a plurality of sample holders at a position on the perimeter of the carousel.
 18. The system of claim 17, wherein the plurality of pneumatic connectors are arranged to enable connection of said one or a plurality of sample holders at equally spaced positions along the perimeter of the carousel.
 19. The system of claim 16, wherein the plurality of stations comprises at least one docking station onto which a matrix droplet extruder is attachable, the docking station comprising a plurality of docking station connectors for connecting to a plurality of docking pneumatic connectors of the matrix droplet extruder, the docking station being controllable by the controller to selectively apply pressure to each of the docking station connectors to control extrusion of liquid droplets from one or more reagent containers of the matrix droplet extruder onto a droplet matrix extrusion surface of the matrix droplet extruder to form the plurality of reagent droplets.
 20. The system of claim 19, wherein said at least one docking station comprises a mechanism for emptying a canister of a reagent into a reagent container of said one or more reagent containers.
 21. The system of claim 19, further comprising a mechanical system that is controlled by the controller to manipulate the substrate of each of the one or plurality of sample holders to place the substrate in contact with the droplet matrix extrusion surface to enable extrusion of the plurality of reagent dots onto the substrate, and to expose the substrate to the analysis module.
 22. The system of claim 21, wherein the mechanical system is controlled by pneumatic pressure.
 23. The system of claim 21, wherein the mechanical system comprises a rinsing chamber that is configured to be placed against the droplet matrix extrusion surface to enable cleaning of the droplet matrix extrusion surface.
 24. The system of claim 15, wherein the analysis module comprises an optical system for acquiring an image of the substrate or an electronic system to measure an electrical property of the substrate.
 25. The system of claim 15, wherein the substrate includes said dots of one or more reagents which are pre-extruded onto the substrate.
 26. The system of claim 15 further including a matrix droplet extruder for printing the plurality of dots of one or more reagents, the matrix droplet extruder comprising: a casing; one or a plurality of reagent containers; a second plurality of pneumatic connectors on the casing that are each configured to be connected to the pneumatic actuator to provide controlled pneumatic pressure to one or more of said second plurality of the pneumatic connectors; a droplet matrix extrusion surface with one or a plurality of printing zones, each printing zone comprising an array of perforations; and a liquid management chip for dispensing one or more reagents from said one or a plurality of reagent containers through the array of perforations of said one or a plurality of printing zones, so as to repeatedly generate a matrix of droplets when applying the pneumatic pressure to said one or more reagents.
 27. The system of claim 26, wherein said one or a plurality of printing zones comprises two printing zones.
 28. The system of claim 26, further comprising a transfer mechanism for transferring each of said one or a plurality of sample holders to and from a position facing the matrix droplet extruder, and for placing the sample slide of each of said one or a plurality of sample holders onto either of said one or a plurality of printing zones.
 29. The system of claim 15, further comprising a disposal container for disposing used sample holders of said one or a plurality of sample holders. 